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Liquid Handling

Justin Beroz, A John Hart
The handheld micropipette is the most ubiquitous instrument for precision handling of microliter-milliliter liquid volumes, which is an essential capability in biology and chemistry laboratories. The range of one pipette is typically adjustable up to 10-fold its minimum volume, requiring the use and maintenance of multiple pipettes for liquid handling across larger ranges. Here we propose a design for a single handheld pipette adjustable from 0.1 μl to 1000 μl (i.e., 10(4)-fold) which spans the range of an entire suite of current commercial pipettes...
November 2016: Review of Scientific Instruments
J C Neubauer, I Sébastien, A Germann, S C Müller, A Meyerhans, H von Briesen, H Zimmermann
The ELISpot assay is used for the detection of T cell responses in clinical trials and vaccine evaluations. Standardization and reproducibility are necessary to compare the results worldwide, inter- and intra-assay variability being critical factors. To assure operator safety as well as high-quality experiment performance, the ELISpot assay was implemented on an automated liquid handling platform, a Tecan Freedom EVO. After validation of the liquid handling, automated loading of plates with cells and reagents was investigated...
November 28, 2016: Cytotechnology
Miguel Baena, Gemma Sangüesa, Natalia Hutter, José María Beltrán, Rosa María Sánchez, Núria Roglans, Marta Alegret, Juan Carlos Laguna
BACKGROUND/OBJECTIVES: Liquid fructose associates with prevalence of type 2 diabetes mellitus and obesity. Intervention studies suggest that metabolically unfit individuals are more responsive than healthy individuals to liquid fructose. We determined whether mice consuming an obesogenic Western diet were more responsive than chow-fed mice to the alterations induced by liquid fructose supplementation (LFS). METHODS: C57BL/6N mice were fed chow or Western diet±ad libitum 15% fructose solution for 12 weeks...
November 2, 2016: Journal of Nutritional Biochemistry
Eduardo Luis de Souza Cruz, Ana Karla da Silva Tabosa, Aline Semblano Carreira Falcão, Talita Tartari, Lucas Machado de Menezes, Edmar Tavares da Costa, José Thiers Carneiro Júnior
PURPOSE: Keratocystic odontogenic tumor (KCOT) is an aggressive benign tumor and the management by complete enucleation followed by cryotherapy maintains the inorganic bone matrix, resulting in better repair and reduces the rates of recurrence. A refrigerant spray with a propane/butane/isobutane gas mixture has been pointed to as an alternative to liquid nitrogen, because the device is easy to handle and contain within the cavity, providing better control and lower risk of injury to the adjacent soft tissue...
November 21, 2016: Oral and Maxillofacial Surgery
Eric Zluhan, Kathleen Kelly, Nick LeClair, Danique Wortel, Kesley Moody
Historically, the routine use of laboratory automation solutions has been prohibitively expensive for many laboratories. As legacy hardware has begun to emerge on the secondary market, automation is becoming an increasingly affordable option to augment workflow in virtually any laboratory. To assess the utility of legacy liquid handling in stem cell differentiation, a used liquid handling robot was purchased at auction to automate a stem cell differentiation protocol that gives rise to CD14 + CD45+ mononuclear cells...
2016: MethodsX
Cosima Koch, Andreas E Posch, Christoph Herwig, Bernhard Lendl
The performance of a fiber optic and an optical conduit in-line attenuated total reflection mid-infrared (IR) probe during in situ monitoring of Penicillium chrysogenum fermentation were compared. The fiber optic probe was connected to a sealed, portable, Fourier transform infrared (FT-IR) process spectrometer via a plug-and-play interface. The optical conduit, on the other hand, was connected to a FT-IR process spectrometer via a knuckled probe with mirrors that had to be adjusted prior to each fermentation, which were purged with dry air...
November 18, 2016: Applied Spectroscopy
Christopher A Hone, Dominique M Roberge, C Oliver Kappe
Molecular oxygen is arguably the greenest reagent available to the organic chemist. Most commonly, a diluted form of oxygen gas, consisting of less than 10 % O2 in N2 ("synthetic air"), is used in pharmaceutical and fine chemical batch manufacturing to effectively address safety concerns when handling molecular oxygen. Concentrations of O2 in N2 below 10 % are generally required to prevent the risk of combustions in the presence of flammable organic solvents ("limiting oxygen concentration"). Nonetheless, the use of pure oxygen is more efficient than using O2 diluted with N2 and can often provide enhanced reaction rates, resulting in significant improvements in product quality and process efficiency...
November 16, 2016: ChemSusChem
Pavel Klimeš, Pavel Mazura, Dušan Turek, Břetislav Brzobohatý
Enzyme kinetic measurements are important for the characterization and engineering of biocatalysts, with applications in a wide range of research fields. The measurement of initial reaction velocity is usually slow and laborious, which motivated us to explore the possibilities for automating this process. Our model enzyme is the maize β-glucosidase Zm-p60.1. Zm-p60.1 plays a significant role in plant growth and development by regulating levels of the active plant hormone cytokinin. Zm-p60.1 belongs to a wide group of hydrolytic enzymes...
November 12, 2016: Protein Science: a Publication of the Protein Society
Miljan Kuljanin, Dylan Z Dieters-Castator, David A Hess, Lynne-Marie Postovit, Gilles A Lajoie
Numerous workflows exist for large scale bottom-up proteomics, many of which achieve exceptional proteome depth. Herein, we evaluated the performance of several commonly used sample preparation techniques for proteomic characterization of HeLa lysates (unfractionated in-solution digests, SDS-PAGE coupled with in-gel digestion, gel-eluted liquid fraction entrapment electrophoresis (GELFrEE) technology, SCX StageTips and High-/Low-pH reversed phase fractionation (HpH)). HpH fractionation was found to be superior in terms of proteome depth (>8400 proteins detected) and fractionation efficiency compared to other techniques...
November 9, 2016: Proteomics
Liu Yang, Rana Said, Mohamed Abdel-Rehim
Microextraction by packed sorbent (MEPS) is a miniaturized form of the solid-phase extraction, is a green sample pretreatment technology. This green sample pretreatment technology has been widely accepted and used by several research groups online or offline as a sample preparation technique before instrument analysis. MEPS reduces the sample volume size and can easily be connected with different chromatographic techniques without modification. The sorbent bed in MEPS is integrated into a liquid handling syringe that allows for low void volume sample manipulations either manually or in combination with laboratory robotics...
November 5, 2016: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Debra D Kristensen, Kate Bartholomew, Shirley Villadiego, Kristina Lorenson
This study attempts to capture the opinions of stakeholders working in immunization programs in low- and middle-income countries to understand how vaccine products could be improved to better meet their needs and to obtain feedback on specific vaccine product attributes including the number of doses per container and ease of preparing a dose for administration. We also reviewed how procurement decisions are made within immunization programs. Semi-structured interviews were undertaken with 158 immunization stakeholders in Brazil, China, India, Peru, the Philippines, and Tanzania...
December 7, 2016: Vaccine
Mohamed A Salem, Jessica Jüppner, Krzysztof Bajdzienko, Patrick Giavalisco
BACKGROUND: The elucidation of complex biological systems requires integration of multiple molecular parameters. Accordingly, high throughput methods like transcriptomics, proteomics, metabolomics and lipidomics have emerged to provide the tools for successful system-wide investigations. Unfortunately, optimized analysis of different compounds requires specific extraction procedures in combination with specific analytical instrumentation. However, the most efficient extraction protocols often only cover a restricted number of compounds due to the different physico-chemical properties of these biological compounds...
2016: Plant Methods
Fidele Tugizimana, Paul A Steenkamp, Lizelle A Piater, Ian A Dubery
Untargeted metabolomic studies generate information-rich, high-dimensional, and complex datasets that remain challenging to handle and fully exploit. Despite the remarkable progress in the development of tools and algorithms, the "exhaustive" extraction of information from these metabolomic datasets is still a non-trivial undertaking. A conversation on data mining strategies for a maximal information extraction from metabolomic data is needed. Using a liquid chromatography-mass spectrometry (LC-MS)-based untargeted metabolomic dataset, this study explored the influence of collection parameters in the data pre-processing step, scaling and data transformation on the statistical models generated, and feature selection, thereafter...
November 3, 2016: Metabolites
Yvonne Herrmann, Andreas Kulawik, Katja Kühbach, Maren Hülsemann, Luriano Peters, Tuyen Bujnicki, Kateryna Kravchenko, Christina Linnartz, Johannes Willbold, Christian Zafiu, Oliver Bannach, Dieter Willbold
OBJECTIVES: Alzheimer's disease (AD) is a neurodegenerative disorder with yet non-existent therapeutic and limited diagnostic options. Reliable biomarker-based AD diagnostics are of utmost importance for the development and application of therapeutic substances. We have previously introduced a platform technology designated 'sFIDA' for the quantitation of amyloid β peptide (Aβ) aggregates as AD biomarker. In this study we implemented the sFIDA assay on an automated platform to enhance robustness and performance of the assay...
November 5, 2016: Clinical Biochemistry
Beth A Rowan, Danelle K Seymour, Eunyoung Chae, Derek S Lundberg, Detlef Weigel
A major goal for biologists is to understand the connection between genes and phenotypic traits, and genetic mapping in experimental populations remains a powerful approach for discovering the causal genes underlying phenotypes. For genetic mapping, the process of genotyping was previously a major rate-limiting step. Modern sequencing technology has greatly improved the resolution and speed of genetic mapping by reducing the time, labor, and cost per genotyping marker. In addition, the ability to perform genotyping-by-sequencing (GBS) has facilitated large-scale population genetic analyses by providing a simpler way to survey segregating genetic variation in natural populations...
2017: Methods in Molecular Biology
Karthik Balajee Laksham, Vinothkumar Kalidoss, Parthibane Sivanantham, Perumal Murthy Sambath, Manoj Kumar Arunachalam, Palanivel Chinnakali
BACKGROUND: Household pesticides (biocides) are widely used for the purpose of controlling vector-borne diseases. Exposure to biocides causes various health hazards in human beings. Prevalence of biocide use, storage methods, and personal protection measures while handling biocides are not known among rural households in South India. MATERIAL AND METHODS: A communitybased cross-sectional study was conducted among 416 households in rural Puducherry (union territory)...
2016: Medycyna Pracy
Hao Jiang, Othonas A Moultos, Ioannis George Economou, Athanassios Z Panagiotopoulos
A polarizable intermolecular potential model with short-range directional hydrogen bonding interactions was developed for water. The model has a rigid geometry with bond lengths and angle set to the experimental gas-phase values. Dispersion interactions are represented by the Buckingham potential assigned to the oxygen atom, while electrostatic interactions are modeled by Gaussian charges. Polarization is handled by a Drude oscillator site, using a negative Gaussian charge attached to the oxygen atom by a harmonic spring...
November 3, 2016: Journal of Physical Chemistry. B
Elaine Mardis, W Richard McCombie
This protocol describes the preparation of plasmid DNA from bacterial cultures or from archived cultures, based on the standard miniprep method. The resulting DNA is suitable in quantity and quality for use as template in capillary DNA sequencing. The protocol relies on the use of the Biomek automated liquid handling system.
November 1, 2016: Cold Spring Harbor Protocols
E Kuijpers, C Bekker, D Brouwer, M Le Feber, W Fransman
Exposure assessment for nano-objects, and their aggregates and agglomerates (NOAA) has evolved from explorative research towards more comprehensive exposure assessment, providing data to further develop currently used conservative control banding (CB) tools for risk assessment. This study aims to provide an overview of current knowledge on emission potential of NOAA across the occupational life cycle stages by a systematic review and subsequently use the results in a data analysis. Relevant parameters that influence emission were collected from peer-reviewed literature with a focus on the four source domains (SD) in the source-receptor conceptual framework for NOAA...
November 1, 2016: Journal of Occupational and Environmental Hygiene
Pei-Han Liu, Pawel L Urban
Dilution is one of the common chemical procedures, which are carried out in all chemistry laboratories - to prepare standard solutions with different concentrations for assay calibration, and to reduce matrix effects while handling complex samples. Yet dilution is mostly performed manually using large-volume manual liquid-handling tools (volumetric flasks, graduated cylinders, and pipettes). Here we describe a simple continuous and automated method of diluting complex samples and stock solutions using an Arduino-based control unit...
November 1, 2016: Analytical Chemistry
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